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Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
143
Original Research Article http://dx.doi.org/10.20546/ijcmas.2017.601.018
Production, Extraction and Characterization of Red Pigment Produced by
Serratia rubidaea JCM 1240T isolated from Soil
R.R. Rakh1*, S.M. Dalvi
2, B.B. Musle
3 and L.S. Raut
4
1Department of Microbiology, Shri Guru Buddhiswami Mahavidyalaya,
Purna (Jn.) Dist. Parbhani, India 2Department of Botany, Shri Guru Buddhiswami Mahavidyalaya,
Purna (Jn.) Dist. Parbhani, India 3Department of Chemistry, Shri Guru Buddhiswami Mahavidyalaya,
Purna (Jn.) Dist. Parbhani, India 4Department of Microbiology, Sant Tukaram College, Parbhani, Dist. Parbhani, India
*Corresponding author
A B S T R A C T
Introduction
Many artificial synthetic colorants, which
have widely been used in foodstuff, dyestuff,
cosmetic and pharmaceutical manufacturing
processes, comprise various hazardous
effects. To counter act the ill effect of
synthetic colorants, there is worldwide
interest in process of development for the
production of pigments from natural sources.
The utilization of natural pigments in
foodstuff, dyestuff, cosmetic and
pharmaceutical manufacturing processes has
been increasing in recent years thanks to the
knowledge about the harmful effects of
synthetic pigments and their industrial
byproducts on humans and the environment
(Unagul et al., 2005; Chidambaram and
Lakshmanaperumalsamy, 2009).
Natural pigments can be obtained from two
major sources, plants (Mizukami, et al., 1978;
Papageorgiou et al., 1979) and micro-
organisms (Cross and Edinberry, 1972; Ryu et
International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 1 (2017) pp. 143-154 Journal homepage: http://www.ijcmas.com
K e y w o r d s
Serratia rubidaea
JCM 1204T,
Prodigiosin,
Presumptive color
test, IR, NMR and
Mass
spectrophotometer.
Accepted:
12 December 2016
Available Online:
10 January 2017
Article Info
A red pigment producing bacteria was isolated from soil sample which was identified by
16S r RNA sequencing as Serratia rubidaea JCM 1204T. The red pigment produced was
tested for prodigiosin by presumptive color test, adsorption spectra, and its quantification
was carried out on King’s B medium. For presumptive color test, the extracted pigment
solution was divided into two portions. One part was acidified with a drop of concentrated
HCl and the other part was alkalinized with a drop of concentrated ammonia solution. A
red or pink color in the acidified solution and a yellow or tan color in the alkaline solution
indicated a positive, presumptive test for prodigiosin. For adsorption spectra, the ethanolic
acidified and alkalinized red pigment extract was tested by spectrophotometer in range
between 300 to 700 nm and found that the ethanolic acidified red pigment extract absorbed
maximum light at 535 nm while ethanolic alkalinized red pigment extract absorbed
maximum light at 470 nm which is characteristic of prodigiosin. The solid red pigment
was also further characterized by IR, NMR, and Mass spectrophotometer, which confirms
that the red pigment produced by Serratia rubidaea JCM 1204T as prodigiosin.
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
144
al., 1989; Parisot et al., 1990; Yongsmith et
al., 1994; Kim et al., 1998a, and Cho et al.,
2002). Biopigments from the micro-
organisms have been preferred over those
from plants because of their stability
(Raisainen et al., 2002) and the availability of
their cultivation technology (Kim et al., 1999;
Parekh et al., 2000) throughout the year. On
the other hand, biopigments from plants have
numerous drawbacks like instability against
light, heat or adverse pH, low water solubility
and often non-availability throughout the year
(Chandni et al., 2012).
One of the studied biopigments of microbial
origin is the prodigiosin. Prodigiosin (PDG) is
a natural red pigment characterized by a
common pyrrolylpyrromethane skeleton,
having low molecular weight (323.4 Dalton),
appearing only in the late stages of bacterial
growth as secondary metabolite (Harris et al.,
2004) and produced by many bacterial
species, including many strains of Serratia
marcescens, Hahella chejuensis,
Streptomyces variegatus, Colwellia (Vibrio)
psychrerythraea, “Pseudomonas
magnesiorubera, and other eubacteria
(Yamamoto et al., 1999). The prodiginine
group, of which prodigiosin is a member, is a
group of structural isomers that contain a
tripyrrole core with different alkyl chains,
have no defined role in the physiology of
producing strains but have been reported to
have antifungal, antibacterial, algicidal,
antiprotozoal, antimalarial, immuno-
suppressive, anticancer and antiproliferative
activities (Castro, 1967; Boger and Patel,
1988;Williams and Quadri, 1980; Demain,
1995; Han et al., 1998;Cerdeno et al., 2001;
Furstner, 2003; Montaner and Pe´rez-Toma´s
2003and Samrot et al., 2011).
According to a recent report, prodigiosin or
its analogues have been considered effective
biological control agents against harmful
algae (Jeong et al., 2005; Nakashima et al.,
2006) have been considered cell growth
regulators (Haddix et al., 2008) and can be
used as a natural dye (Alihosseini, et al.,
2008).
The present investigation focuses on i)
Identification of red pigment producing
bacteria by 16 S rRNA Sequencing ii) Test
the red pigment produced by the red bacteria
for prodigiosin by presumptive color test and
adsorption spectra, and iii) confirmation red
pigment as prodigiosin by IR, NMR and Mass
spectrophotometer.
Materials and Methods
Isolation of Red Pigment Producing
bacteria from Soil
Soil samples, for isolation of red pigment
producing bacteria, were collected from
different field and brought in poly-ethylene
bags to the laboratory, Department of
Microbiology, Shri Guru Buddhiswami
Mahavidyalaya, Purna.
The collected soil samples were serially
diluted and plated on King’s B agar plates.
Red-pigmented colonies were isolated on the
plates after incubation and preserved at 4°C.
The isolated red pigmented bacteria were
identified by 16S r RNA gene sequence
analysis carried out commercially from
Agharkar Research Institute, Pune.
Presumptive color tests for prodigiosin
Red pigment produced in suitable medium
was presumptive tested (Gerber and
Lechevalier, 1976) for prodigiosin. For
presumptive test, pigmented broth was
suspended overnight in 95% ethanol at 22 to
24°C. Debris was removed from the
suspension by centrifugation at 5,000 rpm for
15 min. The clear solution was then divided
into two portions. One part was acidified with
a drop of concentrated HCl and the other part
was alkalinized with a drop of concentrated
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
145
ammonia solution. A red or pink color in the
acidified solution and a yellow or tan color in
the alkaline solution indicative of a positive
presumptive test for prodigiosin.
Determination of adsorption spectra
For determination of absorption spectra,
Gerber and Lechevalier, (1976) method was
used. Pigmented bacteria were treated with
ethanol, and the cell debris in the suspensions
was removed by centrifugation at 5,000 rpm
for 15 min.
Samples (1 ml) of the ethanol extracts were
acidified or alkalinized with 0.1 ml of 1 N
HCI or 1 N NaOH, respectively. The
absorbance spectrum of sample was measured
with a Single Beam UV – Visible
Spectrophotometer (Bioera Life Science Pvt.
Ltd.) in the range of 300 to 700 nm. Acidified
or alkalinized 95% ethanol was used as a
blank.
Production of Prodigiosin in suitable
Medium and Quantification of prodigiosin
In order to determine the suitable media
supporting the maximal production of
prodigiosin, red pigment producing bacteria
was grown in Nutrient Broth, and King’s B
Broth, at 25 °C.The levels of prodigiosin in
these conditions were estimated after 24hr,
48hr, 72hr and 96 hr.
For quantification of prodigiosin produced,
bacterial cell absorbance in the King’s broth
and Nutrient broth was measured at 620 nm,
following which the broth suspensions were
subjected to centrifugation at 5000 rpm for 15
min to collect the cell pellet. 10 ml of 95%
methanol was added to the cell pellet and
centrifuged under the same condition. 95%
methanol was used as a blank. Methanolic
extract of prodigiosin showed characteristic
maxima at 499 nm. Extracted prodigiosin
was estimated using the following equation
(Mekhael and Yousif, 2009).
[OD499 – (1.381 x OD620)] x 1000
Prodigiosin unit/cell =
OD620
Where,
OD499 – pigment absorbance;
OD620 – bacterial cell absorbance
1.381 – constant
Extraction and Characterization of Red
pigment
Serratia rubidaeagrown in King’s B broth
was centrifuged at 10000 rpm for 20 minutes
and the cell pellet was collected. The pigment
from the cell pellet was extracted with
acidified methanol (1N HCl 1ml: Methanol
24 ml) and the extract was centrifuged at
5000 rpm for 15 minutes. The supernatant
was concentrated in Rota evaporator at 60 °C
to obtain crude pigment. The crude
concentrated pigment was recrystallized with
methanol (Min – Jung – Song et al., 2005;
Chandrashekhar et al., 2012).
The chemical structure of the purified
pigment was characterized by FTIR
BRUKER model RXI Spectrometer, mass
spectrometry and NMR (Bruker, Germany).
IR Spectra of prodigiosin was scanned on
FTIR BRUKER model RXI Spectrometer.
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
146
1HNMR spectra were recorded in DMSO on
Gemini 200 MHz instrument using TMS as an
internal standard (chemical shift given in δ
ppm). The mass spectrum was recorded on
Cintra – 15 – GCMS – Spectrophotometer.
Result and Discussion
Isolation of Red Pigment producing
bacteria from Soil
Serially diluted soil samples plated on king’s
B agar plates produced wide range of red
pigmented colony. Out of these colonies, a
red pigment colony was selected whose
pigment was to be tested for prodigiosin,
isolated on the plates after incubation and
preserved at 4°C. Later on the isolated red
pigmented bacterium was identified by 16S r
RNA gene sequence analysis as Serratia
rubidaea JCM 1240T (photo plate 3.1) whose
phylogenetic relation (Figure 3.2) and partial
16S r RNA gene sequence is as follows
(Figure 3.1).
Presumptive color tests for prodigiosin
For this presumptive test pigmented broth was
suspended overnight in 95% ethanol at 22 to
24°C. Debris was removed from the
suspension by centrifugation at 5,000 rpm for
15 min. The clear solution was then divided
into two portions. One part was acidified with
a drop of concentrated HCl and the other part
was alkalinized with a drop of concentrated
ammonia solution. A red or pink color in the
acidified solution was observed while a
yellow or tan color in the alkaline solution
indicated a positive presumptive test for
prodigiosin as shown (photo plate 2).
Similar types of results were also shown in
studies carried out by Ming-Jer Ding and
Robert P. Williams (1983) where strain of
Serratia marcescens isolated from clinic
shows a red or pink color in the acidified
solution while a yellow or tan color in the
alkaline solution. This test indicative for
presumptive test for prodigiosin.
Determination of adsorption spectra
For determination of absorption spectra,
pigmented ethanol extracts were acidified or
alkalinized with 0.1 ml of 1 N HCI or 1 N
NaOH, respectively. The absorbance
spectrum of sample was measured with a
Single Beam UV – Visible Spectrophotometer
(Bioera Life Science Pvt. Ltd.) in the range of
300 to 700 nm. Acidified or alkalinized 95%
ethanol was used as a blank.
Pigment extracted with ethanol from Serratia
rubidaea JCM 1240T showed characteristic
maxima of 535 (Figure 3.3) and 470 nm in
acid and alkaline solutions, respectively,
which is indicative that the pigment extracted
with ethanol was prodigiosin. Similarly,
Prodigiosin extracted with ethanol from S.
marcescens Nima showed characteristic
maxima of 535 and 470 nm in acid and
alkaline solutions, respectively (Williams and
Qadri, 1980; Ming-Jer Ding and Robert
Williams, 1983).
Prodigiosin was reported to display a
characteristic absorption spectrum in ethanol,
with a maximum at 534 nm (Slater et al.,
2003) and single peak absorbance at 535 Nm
(Giri et al., 2004; Song et al., 2006; Montaner
et al., 2000) extracted prodigiosin by shaking
the S. marcescens 2170 cells with a mixture
of methanol/lN HCl in the ratio 24:l.
Absorption spectrum of the pigment produced
by Serratia rubidaea JCM was dependent on
pH value similar to that reported earlier (Song
et al., 2006). Thus at pH 2.0, the pigment was
red and showed a maximum absorption at 535
nm, which is identical to that of prodigiosin
hydrochloride. Under neutral condition (pH
7.3), its absorption intensity decreased and the
colour of the pigment changed to pink.
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
147
However, in alkaline condition (pH 10.0) the
colour was orange and its absorption
spectrum shifted to 470 nm. Pigment
produced by Serratia rubidaea JCM 1240T
recorded maximum absorption at and 470 nm
in acid and alkaline solutions, respectively
suggesting that this pigment is prodigiosin.
Production of Prodigiosin in suitable
Medium and Quantification of prodigiosin
To determine suitable medium for production
and quantification of prodigiosin,Serratia
rubidaea JCM 1240T was grown in Nutrient
Broth, King’s B Broth, at 25 °C.
For quantification of prodigiosin produced,
bacterial cell absorbance in the medium was
measured at 620 nm, while absorbance of
methanolic extract of prodigiosin was
measured at 499 nm (Table 3.1). The
concentrations of prodigiosin in these
conditions were estimated after 0hr, 24hr,
48hr 72hr and 96 hr. By using these
absorbance, prodigiosin concentration was
determined as represented in (Table 3.2).
Table.3.1 Pigment Absorbance at 499nm
Hour Nutrient Broth (O.D.) King’s B Broth (O.D.)
24 hr 00 00
48 hr 1.603 1.673 72 hr 1.654 1.686 96 hr 1.566 1.580
Table.3.2 Total prodigiosin estimated in nutrient broth and King’s B broth
Hour Prodigiosin in
Nutrient Broth mg/lit
Prodigiosin in
King’s B Broth mg/ lit
24 hr 00 00
48 hr 1070.33 1188.94
72 hr 1055.96 1246.10
96 hr 885.38 957.10
Fig 3.1
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
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Fig 3.2
Fig 3.3
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149
Fig 3.4
Fig 3.5
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
150
Photo 3.1
Photo 3.2
Prodigiosin unit per cell after 48hr was
found to be 1070.33 and 1188.94 in
nutrient broth and King’s B broth
respectively. Maximum production was
reported in 72hr thereafter production ceased
towards 96hr. The lowest prodigiosin unit per
cell was found after 96hr i.e. in nutrient broth
it was 885 and in peptone glycerol broth i.e.
957.1.
Similar results were obtained by Kamble and
Hiwarale (2012) where they compared
prodigiosin production in Nutrient Broth and
Peptone Glycerol Broth. In this study, they
found highest prodigiosin production after 72
hr in peptone glycerol broth than nutrient
broth.
Extraction and Characterization of Red
pigment
The chemical structure of the purified
prodigiosin was characterized by FTIR
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
151
BRUKER model RXI Spectrometer, mass
spectrometry and NMR (Bruker, Germany).
IR Spectra of prodigiosin was scanned on
FTIR BRUKER model RXI Spectrometer. 1HNMR spectra were recorded in DMSO on
Gemini 200 MHz instrument using TMS as an
internal standard (chemical shift given in δ
ppm). The mass spectrum was recorded on
Cintra – 15 – GCMS – Spectrophotometer.
IR Spectra
IR Spectra of prodigiosin (Figure 3.4) showed
characteristic band at near region 3442.14 cm-
1 due to group (amide gr.), bands at
2924.16 – 2853.48 cm-1
due to C – H (sp3)
stretching vibrations from methyl and
Methylene group, bands at near region
1633.96 cm-1
is due to Pyrrole group and the
band at 1716.73 cm-1
is due to
stretching vibrations. The visible peak at
1403.46 cm-1
is due to the presence of C–O
group in prodigiosin.
Thus from the IR Spectrum the main
functional groups that resulted in red pigment
were Pyrrole, amide, Methylene, methyl and
alkene.
1HNMR spectra: 1HNMR spectra of prodigiosin were studied
in DMSO, represents the proton peaks
corresponding to chemical shifts at 𝛿5.323
ppm, 𝛿5.053 ppm, 𝛿3.773 ppm, 𝛿2.354 ppm,
𝛿1.236ppm, and 𝛿0.853 ppm assigned to the
carbon atoms C2, C12, C11, C18, C21, and
C22 based on the 1HNMR spectra structure of
prodigiosin presented (Figure 3.5).
Mass Spectra
The mass spectra for the prodigiosin exhibited
correct molecular ion peaks for its molecular
weights.
In the present study, the data obtained for the
spectroscopic analyses of the red pigment
with IR, 1HNMR and Mass spectra very
clearly testify that the pigment produced by
Serratia rubidaea JCM 1240T is prodigiosin-
like pigment. Sumathi et al., 2014 used same
technique to characterize the pigment
produced bySerratia marcescens NPLR1
where they found the pigment produced was
also prodigiosin.
References
Alihosseini, F., K.S. Ju, J. Lango, B.D.
Hammock, and G. Sun. 2008.
Antibacterial colorants: characterization
of prodiginines and their applications on
textile materials. Biotechnol. Prog., 24:
742–747.
Boger, D.L., and M. Patel. 1988. Total
synthesis of prodigiosin, prodigiosene,
and desmethoxyprodigiosin: Diels-
Alder reactions of heterocyclic
azadienes and development of an
effective palladium(II)-promoted 2,2_-
bipyrrole coupling procedure. J. Org.
Chem., 53: 1405–1415.
Castro, A.J. 1967. Antimalarial activity of
prodigiosin. Nature, 213: 903–904.
Cerdeno, A.M., Bibb, M.J. and Challis, G.L.
2001. Analysis of the prodiginine
biosynthesis gene cluster of
Streptomyces coelicolorA3(2): new
mechanisms for chain initiation and
termination in modular multienzymes.
Chem. Biol., 8, 817-829.
Chandni Gulani, Sourav Bhattacharya and
Arijit Das. 2012. Assessment of process
parameters influencing the enhanced
production of prodigiosin from Serratia
marcescens and evaluation of its
antimicrobial, antioxidant and dyeing
potentials. Malaysian J. Microbiol.,
Vol. 8(2), pp. 116-122
Chandrashekhar Naik, Srisevita, J.M.,
Shusham, K.N., Farah Noorin, Shilpa,
A.C., Muttanna, C.D., Darshan, N.,
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
152
Sannadurgappa, D. 2012. Peanut Oil
Cake: A Novel Substrate for Enhanced
Cell Growth and Prodigiosin Production
from Serratia marcescens CF – 53. J.
Res. Biol., 2 (6) : 549 – 557.
Chidambaram Kulandaisamy Venil and
Perumalsamy Lakshmanaperumalsamy.
2009. An Insightful Overview on
Microbial Pigment, Prodigiosin.
Electronic J. Biol., Vol. 5(3): 49-61.
Cho, Y.J., Park J.P., Hwang H.J., Kim, S.W.,
Choi, J.W., and Yun, J.W. 2002.
Production of red pigment by
submerged culture of Paecilomyces
sinclairii. Lett. Appl. Microbiol., 35:
195-202.
Cross, B.E., and Edinberry, M.N. 1972.
Pigments of Gnomonia erythrostoma.
Part I. The structures of
erythrostominone,
deoxyerythrostominone, and
deoxyerythrostominol. J. Chem. Soc.
Perkin I, 3: 380-390.
Demain, A.L. 1995. Why do microorganisms
produce antimicrobials? In Fifty Years
of Antimicrobials: Past Perspectives and
Future Trends (Society for General
Microbiology Symposium no. 53), pp.
205–228. Edited by P. A. Hunter, G. K.
Darby & N. J. Russell. Cambridge:
Cambridge University Press.
Furstner, A. 2003. Chemistry and biology of
roseophilin and the Prodigiosin
alkaloids: A survey of the last 2500
years. Chem. Int. Ed. Engl., 42, 3582-
3603.
Gerber, N.N. and Lechevalier, M.P. 1976.
Prodiginine (prodigiosin-like) pigments
from Streptomyces and other
Actinomyces. Canadian J. Microbiol.,
22: 658-667.
Giri, A.V., Anandkumar, N., Muthukumaran,
G., Pennathur, G. 2004. A novel
medium for the enhanced cell growth
and production of prodigiosin from
Serratia marcescens isolated from soil.
Biomedical Central Microbiol., 4: 1-10.
Haddix, P.L., et al. 2008. Kinetic analysis of
growth rate, ATP, and pigmentation
suggests an energy-spilling function for
the pigment prodigiosin of Serratia
marcescens. J. Bacteriol., 190: 7453–
7463.
Han, S.B., et al. 1998. T-cell specific
immunosuppression by prodigiosin
isolated from Serratia marcescens. Int.
J. Immunopharmacol., 20: 1–13.
Harris, K.P., Williamson, R., Slater, H., Cox,
A., Abbasi, S., Foulds, I., Simonsen, T.,
Leeper J., and Salmond P.C. 2004. The
Serratia gene cluster encoding
biosynthesis of the red antibiotic,
prodigiosin, shows species and strain
dependent genome context variation.
Microbiol, 150: 3547-3560.
Jeong, H., et al. 2005. Genomic blueprint of
Hahella chejuensis, a marine microbe
producing an algicidal agent. Nucleic
Acids Res., 33:7066–7073.
Kamble, K.D., and Hiwarale, V.D. 2012.
Prodigiosin production from Serratia
marcescens strains obtained from farm
soil, Int. J. Environ. Sci., Volume 3, No
1, pp 631 – 638.
Kim, C.H., Kim, S.W., and Hong, S.I. 1998.
Production of red pigment by Serratia
sp. KH-95 and its cultural properties.
Korean J. Biotechnol. Bioeng., 13: 431-
437.
Kim, C.H., Kim, S.W., and Hong, S.I. 1999.
An integrated fermentation separation
process for the production of red
pigment by Serratia sp. KH-95. Process
Biochem., 35: 485-490.
Mekhael, R., and Yousif, S.Y. 2009. The role
of red pigment produced by Serratia
marcescens as antibacterial and plasmid
curing agent. J. Duhok Univ., 12: 268-
274.
Min – Jung – Song, Jungdon Bae, Dae Sil
Lee, Chnag – Ho Kim, Seung – Wook
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
153
Kim, Suk – In Hong. 2006. purification
and characterization of prodigiosin
produced by integrated bioreactor from
Serratia spp. KH – 95. J. Biosc. Bioeng.,
101 : 157 – 161.
Ming-Jer Ding and Robert P. Williams. 1983.
Biosynthesis of Prodigiosin by White
Strains of Serratia marcescens Isolated
from Patients. J. Clin. Microbiol., vol.
17, No. 3 p. 476 – 480.
Mizukami, H., Konoshima, M., and Tabata,
M. 1978 Variation in pigment
production in Lithospermum
erythrorhizon callus cultures.
Phytochem., 17: 95-97.
Montaner, B., and R. Pe´rez-Toma´s. 2003.
The prodigiosins: a new family of
anticancer drugs. Curr. Cancer Drug
Targets, 3: 57–65.
Montaner, B., Navarro, S., Pique M. Vilaseca
M. Martinell, M., Giralt, E., Gil, J. and
Perez-Thomas, R., 2000. Prodigiosin
from the supernatant of Serratia
marcescens induce apoptosis in
haematopoietic cancer cell lines, British
J. Pharmacol., 131(3), pp 585-593.
Nakashima, T., et al. 2006. Producing
mechanism of an algicidal compound
against red tide phytoplankton in a
marine bacterium -proteobacterium.
Appl. Microbiol. Biotechnol., 73:684–
690.
Papageorgiou V. P., Winkler A., Sagredos A.
N., and Digenis G. A. 1979. Studies on
the relationship of structure to
antimicrobial properties of
naphthoquinones and other constituents
of Alkanna tinctoria. Planta Med., 35:
56-60.
Parekh S., Vinci, V. A. and Strobel, R. J.
2000. Improvement of microbial strains
and fermentation processes. Appl.
Microbiol. Biotechnol., 54: 287-301.
Parisot D., Devys M., and Barbier M. 1990.
Naphthoquinone pigments related to
fusarubin from the fungus Fusarium
solani (Mart.) Sacc. Microbios, 64: 31-
47.
Raisainen, R., Nousiainen, P. and Hynninen,
P.H. 2002. Dermorubin and 5-
chlorodermorubin natural anthraquinone
carboxylic acids as dyes for wool.
Textile Res. J., 72: 973-976.
Ryu B. H., Park B. G., Chi Y. E., and Lee J.
H. 1989. Production of purplish-red
pigment in mixed culture of
Streptomyces propurpuratus ATCC
21630 and Bacillus sp R-89. Korean J
Appl. Microbiol. Bioeng, 17: 327-333.
Samrot, A. V., Chandana, K., Senthilkumar P.
and Narendra, K. G. 2011. Optimization
of Prodigiosin production by Serratia
marcescens SU-10 and evaluation of its
bioactivity. Int. Res. J. Biotechnol.,
(ISSN: 2141-5153), 2(5), 128-133.
Shahitha, S. and K. Poornima. 2012.
Enhanced Production of Prodigiosin
Production in Serratia Marcescens. J.
Appl. Pharmaceutical Sci., 02(08); 138-
140.
Slater, H., Crow, M., Everson, L. and
Salmond, G.P.C. 2003. Phosphate
availability regulates biosynthesis of
two antibiotics, Prodigiosin and
carbapenem, in Serratia via both
quorum-sensing-dependentand –
independent pathways. Mol. Microbiol.,
47(2), 303–320.
Song, M.J., Bae, J., Lee, D.S., Kim, C.H.,
Kim, J.S., Kim, S.W., Hong, S.I. 2006.
Purification and characterization of
prodigiosin produced by integrated
bioreactor from Serratia sp. KH-95. J.
Biosci. Bioeng., 101: 157-161.
Unagul, P., Wongsa, P., Kittakoop, P.,
Intamas, S., Srikiti-Kulchai, P.,
Tanticharoen, M. 2005. Production of
red pigments by the insect pathogenic
fungus Cordyceps unilateralis BCC
1869. J. Ind. Microbiol. Biotechnol., 32:
135-140.
Int.J.Curr.Microbiol.App.Sci (2017) 6(1): 143-154
154
Williams, R.P. and Quadri, S.M. 1980. The
pigments of Serratia. In The Genus
Serratia, pp. 31–75. Edited by A. Von
Graevenitz & S. J. Rubin. Boca Raton,
FL: CRC Press Inc.
Yamamoto, C., et al. 1999. Cycloprodigiosin
hydrochloride, a new H(+)/ Cl(-)
symporter, induces apoptosis in human
and rat hepatocellular cancer cell lines
in vitro and inhibits the growth of
hepatocellular carcinoma xenografts in
nude mice. Hepatol., 30: 894–902.
Yongsmith, B., Krairak, S., and Bavavoda, R.
1994. Production of yellow pigments in
submerged culture of a mutant of
Monascus sp. J. Ferment. Bioeng., 78:
223-228.
How to cite this article:
Rakh, R.R., S.M. Dalvi, B.B. Musle and Raut, L.S. 2017. Production, Extraction and
Characterization of Red Pigment Produced by Serratia rubidaea JCM 1240T isolated from
Soil. Int.J.Curr.Microbiol.App.Sci. 6(1): 143-154.
doi: http://dx.doi.org/10.20546/ijcmas.2017.601.018